CA2016211C - Multiply mutated subtilisins - Google Patents

Multiply mutated subtilisins

Info

Publication number: CA2016211C
Authority: CA; Canada
Prior art keywords: subtilisin; ser; ala; gly; val
Prior art date: 1989-05-17
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.): Expired - Lifetime

Application number

CA002016211A

Other languages

English (en)

French (fr)

Other versions

CA2016211A1 (en

Inventor

Mark M. Zukowski

Linda O. Narhi

Michael Levitt

Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)

Amgen Inc

Original Assignee

Amgen Inc

Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)

1989-05-17

Filing date

1990-05-07

Publication date

1998-07-21

1990-05-07 Application filed by Amgen Inc filed Critical Amgen Inc

1990-11-17 Publication of CA2016211A1 publication Critical patent/CA2016211A1/en

1998-07-21 Application granted granted Critical

1998-07-21 Publication of CA2016211C publication Critical patent/CA2016211C/en

2010-05-07 Anticipated expiration legal-status Critical

Status Expired - Lifetime legal-status Critical Current

Links

108010056079 Subtilisins Proteins 0.000 title claims abstract description 81
102000005158 Subtilisins Human genes 0.000 title claims abstract description 75
108090000787 Subtilisin Proteins 0.000 claims abstract description 204
235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 66
241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 32
239000000203 mixture Substances 0.000 claims abstract description 32
244000063299 Bacillus subtilis Species 0.000 claims abstract description 12
150000001413 amino acids Chemical group 0.000 claims description 53
230000000694 effects Effects 0.000 claims description 40
239000003599 detergent Substances 0.000 claims description 35
238000012986 modification Methods 0.000 claims description 22
230000004048 modification Effects 0.000 claims description 22
KLKHFFMNGWULBN-VKHMYHEASA-N Asn-Gly Chemical group NC(=O)C[C@H](N)C(=O)NCC(O)=O KLKHFFMNGWULBN-VKHMYHEASA-N 0.000 claims description 20
ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Chemical compound CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 claims description 14
238000009472 formulation Methods 0.000 claims description 11
RLMISHABBKUNFO-WHFBIAKZSA-N Ala-Ala-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O RLMISHABBKUNFO-WHFBIAKZSA-N 0.000 claims description 9
FFALDIDGPLUDKV-ZDLURKLDSA-N Gly-Thr-Ser Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O FFALDIDGPLUDKV-ZDLURKLDSA-N 0.000 claims description 3
PVMPDMIKUVNOBD-CIUDSAMLSA-N Leu-Asp-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O PVMPDMIKUVNOBD-CIUDSAMLSA-N 0.000 claims description 3
NVNPWELENFJOHH-CIUDSAMLSA-N Ser-Ser-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CO)N NVNPWELENFJOHH-CIUDSAMLSA-N 0.000 claims description 3
DJDSEDOKJTZBAR-ZDLURKLDSA-N Thr-Gly-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O DJDSEDOKJTZBAR-ZDLURKLDSA-N 0.000 claims description 3
XOWKUMFHEZLKLT-CIQUZCHMSA-N Thr-Ile-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O XOWKUMFHEZLKLT-CIQUZCHMSA-N 0.000 claims description 3
MVBWLRJESQOQTM-ACZMJKKPSA-N Ala-Gln-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O MVBWLRJESQOQTM-ACZMJKKPSA-N 0.000 claims description 2
YIGLXQRFQVWFEY-NRPADANISA-N Ala-Gln-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O YIGLXQRFQVWFEY-NRPADANISA-N 0.000 claims description 2
WUHJHHGYVVJMQE-BJDJZHNGSA-N Ala-Leu-Ile Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WUHJHHGYVVJMQE-BJDJZHNGSA-N 0.000 claims description 2
AOAKQKVICDWCLB-UWJYBYFXSA-N Ala-Tyr-Asn Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N AOAKQKVICDWCLB-UWJYBYFXSA-N 0.000 claims description 2
LYILPUNCKACNGF-NAKRPEOUSA-N Ala-Val-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](C)N LYILPUNCKACNGF-NAKRPEOUSA-N 0.000 claims description 2
XCIGOVDXZULBBV-DCAQKATOSA-N Ala-Val-Lys Chemical compound CC(C)[C@H](NC(=O)[C@H](C)N)C(=O)N[C@@H](CCCCN)C(O)=O XCIGOVDXZULBBV-DCAQKATOSA-N 0.000 claims description 2
REWSWYIDQIELBE-FXQIFTODSA-N Ala-Val-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O REWSWYIDQIELBE-FXQIFTODSA-N 0.000 claims description 2
XVLLUZMFSAYKJV-GUBZILKMSA-N Arg-Asp-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O XVLLUZMFSAYKJV-GUBZILKMSA-N 0.000 claims description 2
GNKVBRYFXYWXAB-WDSKDSINSA-N Asn-Glu-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O GNKVBRYFXYWXAB-WDSKDSINSA-N 0.000 claims description 2
NPZJLGMWMDNQDD-GHCJXIJMSA-N Asn-Ser-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O NPZJLGMWMDNQDD-GHCJXIJMSA-N 0.000 claims description 2
GXIUDSXIUSTSLO-QXEWZRGKSA-N Asp-Val-Met Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CC(=O)O)N GXIUDSXIUSTSLO-QXEWZRGKSA-N 0.000 claims description 2
YJIUYQKQBBQYHZ-ACZMJKKPSA-N Gln-Ala-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O YJIUYQKQBBQYHZ-ACZMJKKPSA-N 0.000 claims description 2
RHRLHXQWHCNJKR-PMVVWTBXSA-N Gly-Thr-His Chemical compound NCC(=O)N[C@@H]([C@H](O)C)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 RHRLHXQWHCNJKR-PMVVWTBXSA-N 0.000 claims description 2
OWYIDJCNRWRSJY-QTKMDUPCSA-N His-Pro-Thr Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O OWYIDJCNRWRSJY-QTKMDUPCSA-N 0.000 claims description 2
241000880493 Leptailurus serval Species 0.000 claims description 2
WQWSMEOYXJTFRU-GUBZILKMSA-N Leu-Glu-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O WQWSMEOYXJTFRU-GUBZILKMSA-N 0.000 claims description 2
OVZLLFONXILPDZ-VOAKCMCISA-N Leu-Lys-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OVZLLFONXILPDZ-VOAKCMCISA-N 0.000 claims description 2
AMSSKPUHBUQBOQ-SRVKXCTJSA-N Leu-Ser-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)O)N AMSSKPUHBUQBOQ-SRVKXCTJSA-N 0.000 claims description 2
HUKLXYYPZWPXCC-KZVJFYERSA-N Met-Ala-Thr Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O HUKLXYYPZWPXCC-KZVJFYERSA-N 0.000 claims description 2
LPGSNRSLPHRNBW-AVGNSLFASA-N Pro-His-Val Chemical compound C([C@@H](C(=O)N[C@@H](C(C)C)C([O-])=O)NC(=O)[C@H]1[NH2+]CCC1)C1=CN=CN1 LPGSNRSLPHRNBW-AVGNSLFASA-N 0.000 claims description 2
BTKUIVBNGBFTTP-WHFBIAKZSA-N Ser-Ala-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)NCC(O)=O BTKUIVBNGBFTTP-WHFBIAKZSA-N 0.000 claims description 2
LALNXSXEYFUUDD-GUBZILKMSA-N Ser-Glu-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O LALNXSXEYFUUDD-GUBZILKMSA-N 0.000 claims description 2
IOVHBRCQOGWAQH-ZKWXMUAHSA-N Ser-Gly-Ile Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(O)=O IOVHBRCQOGWAQH-ZKWXMUAHSA-N 0.000 claims description 2
MQUZANJDFOQOBX-SRVKXCTJSA-N Ser-Phe-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O MQUZANJDFOQOBX-SRVKXCTJSA-N 0.000 claims description 2
XJDMUQCLVSCRSJ-VZFHVOOUSA-N Ser-Thr-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O XJDMUQCLVSCRSJ-VZFHVOOUSA-N 0.000 claims description 2
CAJFZCICSVBOJK-SHGPDSBTSA-N Thr-Ala-Thr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O CAJFZCICSVBOJK-SHGPDSBTSA-N 0.000 claims description 2
HHPSUFUXXBOFQY-AQZXSJQPSA-N Trp-Thr-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N)O HHPSUFUXXBOFQY-AQZXSJQPSA-N 0.000 claims description 2
ASQFIHTXXMFENG-XPUUQOCRSA-N Val-Ala-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O ASQFIHTXXMFENG-XPUUQOCRSA-N 0.000 claims description 2
ZLFHAAGHGQBQQN-GUBZILKMSA-N Val-Ala-Pro Natural products CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O ZLFHAAGHGQBQQN-GUBZILKMSA-N 0.000 claims description 2
COYSIHFOCOMGCF-UHFFFAOYSA-N Val-Arg-Gly Natural products CC(C)C(N)C(=O)NC(C(=O)NCC(O)=O)CCCN=C(N)N COYSIHFOCOMGCF-UHFFFAOYSA-N 0.000 claims description 2
BMGOFDMKDVVGJG-NHCYSSNCSA-N Val-Asp-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N BMGOFDMKDVVGJG-NHCYSSNCSA-N 0.000 claims description 2
108010069490 alanyl-glycyl-seryl-glutamic acid Proteins 0.000 claims description 2
108010027668 glycyl-alanyl-valine Proteins 0.000 claims description 2
108010050848 glycylleucine Proteins 0.000 claims description 2
108010015792 glycyllysine Proteins 0.000 claims description 2
108010078274 isoleucylvaline Proteins 0.000 claims description 2
108010083708 leucyl-aspartyl-valine Proteins 0.000 claims description 2
108010044311 leucyl-glycyl-glycine Proteins 0.000 claims description 2
108010031719 prolyl-serine Proteins 0.000 claims description 2
108010071207 serylmethionine Proteins 0.000 claims description 2
ZVFVBBGVOILKPO-WHFBIAKZSA-N Ala-Gly-Ala Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O ZVFVBBGVOILKPO-WHFBIAKZSA-N 0.000 claims 1
VNYMOTCMNHJGTG-JBDRJPRFSA-N Ala-Ile-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O VNYMOTCMNHJGTG-JBDRJPRFSA-N 0.000 claims 1
HHRAXZAYZFFRAM-CIUDSAMLSA-N Ala-Leu-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O HHRAXZAYZFFRAM-CIUDSAMLSA-N 0.000 claims 1
IPZQNYYAYVRKKK-FXQIFTODSA-N Ala-Pro-Ala Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O IPZQNYYAYVRKKK-FXQIFTODSA-N 0.000 claims 1
XSLGWYYNOSUMRM-ZKWXMUAHSA-N Ala-Val-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O XSLGWYYNOSUMRM-ZKWXMUAHSA-N 0.000 claims 1
VHAQSYHSDKERBS-XPUUQOCRSA-N Ala-Val-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O VHAQSYHSDKERBS-XPUUQOCRSA-N 0.000 claims 1
DXZNJWFECGJCQR-FXQIFTODSA-N Asn-Asn-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC(=O)N)N DXZNJWFECGJCQR-FXQIFTODSA-N 0.000 claims 1
KEUNWIXNKVWCFL-FXQIFTODSA-N Asn-Met-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CO)C(O)=O KEUNWIXNKVWCFL-FXQIFTODSA-N 0.000 claims 1
SUIJFTJDTJKSRK-IHRRRGAJSA-N Asn-Pro-Tyr Chemical compound NC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 SUIJFTJDTJKSRK-IHRRRGAJSA-N 0.000 claims 1
VLDRQOHCMKCXLY-SRVKXCTJSA-N Asn-Ser-Phe Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O VLDRQOHCMKCXLY-SRVKXCTJSA-N 0.000 claims 1
LMIWYCWRJVMAIQ-NHCYSSNCSA-N Asn-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N LMIWYCWRJVMAIQ-NHCYSSNCSA-N 0.000 claims 1
CLUMZOKVGUWUFD-CIUDSAMLSA-N Asp-Leu-Asn Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O CLUMZOKVGUWUFD-CIUDSAMLSA-N 0.000 claims 1
KGHLGJAXYSVNJP-WHFBIAKZSA-N Asp-Ser-Gly Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O KGHLGJAXYSVNJP-WHFBIAKZSA-N 0.000 claims 1
UXRVDHVARNBOIO-QSFUFRPTSA-N Asp-Val-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(=O)O)N UXRVDHVARNBOIO-QSFUFRPTSA-N 0.000 claims 1
RGXXLQWXBFNXTG-CIUDSAMLSA-N Gln-Arg-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O RGXXLQWXBFNXTG-CIUDSAMLSA-N 0.000 claims 1
WQWMZOIPXWSZNE-WDSKDSINSA-N Gln-Asp-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O WQWMZOIPXWSZNE-WDSKDSINSA-N 0.000 claims 1
YXQCLIVLWCKCRS-RYUDHWBXSA-N Gln-Gly-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CCC(=O)N)N)O YXQCLIVLWCKCRS-RYUDHWBXSA-N 0.000 claims 1
MTCXQQINVAFZKW-MNXVOIDGSA-N Gln-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)N)N MTCXQQINVAFZKW-MNXVOIDGSA-N 0.000 claims 1
BYKZWDGMJLNFJY-XKBZYTNZSA-N Gln-Ser-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)N)N)O BYKZWDGMJLNFJY-XKBZYTNZSA-N 0.000 claims 1
LJPIRKICOISLKN-WHFBIAKZSA-N Gly-Ala-Ser Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O LJPIRKICOISLKN-WHFBIAKZSA-N 0.000 claims 1
JLJLBWDKDRYOPA-RYUDHWBXSA-N Gly-Gln-Tyr Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 JLJLBWDKDRYOPA-RYUDHWBXSA-N 0.000 claims 1
HAXARWKYFIIHKD-ZKWXMUAHSA-N Gly-Ile-Ser Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O HAXARWKYFIIHKD-ZKWXMUAHSA-N 0.000 claims 1
GWCJMBNBFYBQCV-XPUUQOCRSA-N Gly-Val-Ala Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O GWCJMBNBFYBQCV-XPUUQOCRSA-N 0.000 claims 1
BAYQNCWLXIDLHX-ONGXEEELSA-N Gly-Val-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)CN BAYQNCWLXIDLHX-ONGXEEELSA-N 0.000 claims 1
UAVQIQOOBXFKRC-BYULHYEWSA-N Ile-Asn-Gly Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O UAVQIQOOBXFKRC-BYULHYEWSA-N 0.000 claims 1
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DCQMJRSOGCYKTR-GHCJXIJMSA-N Ile-Asp-Ser Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O DCQMJRSOGCYKTR-GHCJXIJMSA-N 0.000 claims 1
LEHPJMKVGFPSSP-ZQINRCPSSA-N Ile-Glu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)[C@@H](C)CC)C(O)=O)=CNC2=C1 LEHPJMKVGFPSSP-ZQINRCPSSA-N 0.000 claims 1
UGTHTQWIQKEDEH-BQBZGAKWSA-N L-alanyl-L-prolylglycine zwitterion Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O UGTHTQWIQKEDEH-BQBZGAKWSA-N 0.000 claims 1
RCFDOSNHHZGBOY-UHFFFAOYSA-N L-isoleucyl-L-alanine Natural products CCC(C)C(N)C(=O)NC(C)C(O)=O RCFDOSNHHZGBOY-UHFFFAOYSA-N 0.000 claims 1
VWHGTYCRDRBSFI-ZETCQYMHSA-N Leu-Gly-Gly Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)NCC(O)=O VWHGTYCRDRBSFI-ZETCQYMHSA-N 0.000 claims 1
OYQUOLRTJHWVSQ-SRVKXCTJSA-N Leu-His-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CO)C(O)=O OYQUOLRTJHWVSQ-SRVKXCTJSA-N 0.000 claims 1
UCBPDSYUVAAHCD-UWVGGRQHSA-N Leu-Pro-Gly Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O UCBPDSYUVAAHCD-UWVGGRQHSA-N 0.000 claims 1
GQFDWEDHOQRNLC-QWRGUYRKSA-N Lys-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN GQFDWEDHOQRNLC-QWRGUYRKSA-N 0.000 claims 1
IEIHKHYMBIYQTH-YESZJQIVSA-N Lys-Tyr-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)[C@H](CCCCN)N)C(=O)O IEIHKHYMBIYQTH-YESZJQIVSA-N 0.000 claims 1
KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 claims 1
108010079364 N-glycylalanine Proteins 0.000 claims 1
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Classifications

- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
- C12N9/54—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea bacteria being Bacillus

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Life Sciences & Earth Sciences (AREA)
Chemical & Material Sciences (AREA)
Health & Medical Sciences (AREA)
Genetics & Genomics (AREA)
Organic Chemistry (AREA)
Zoology (AREA)
Engineering & Computer Science (AREA)
Bioinformatics & Cheminformatics (AREA)
Wood Science & Technology (AREA)
Microbiology (AREA)
Biotechnology (AREA)
Biomedical Technology (AREA)
Molecular Biology (AREA)
Biochemistry (AREA)
General Engineering & Computer Science (AREA)
General Health & Medical Sciences (AREA)
Medicinal Chemistry (AREA)
Detergent Compositions (AREA)
Micro-Organisms Or Cultivation Processes Thereof (AREA)
Enzymes And Modification Thereof (AREA)
Cosmetics (AREA)
Tires In General (AREA)

CA002016211A 1989-05-17 1990-05-07 Multiply mutated subtilisins Expired - Lifetime CA2016211C (en)

Applications Claiming Priority (2)

Application Number	Priority Date	Filing Date	Title
US35312489A	1989-05-17	1989-05-17
US353,124		1989-05-17

Publications (2)

Publication Number	Publication Date
CA2016211A1 CA2016211A1 (en)	1990-11-17
CA2016211C true CA2016211C (en)	1998-07-21

Family

ID=23387862

Family Applications (1)

Application Number	Title	Priority Date	Filing Date
CA002016211A Expired - Lifetime CA2016211C (en)	1989-05-17	1990-05-07	Multiply mutated subtilisins

Country Status (13)

Country	Link
US (1)	US5397705A (de)
EP (1)	EP0398539B1 (de)
JP (1)	JPH03505976A (de)
KR (1)	KR100188532B1 (de)
AT (1)	ATE141324T1 (de)
AU (1)	AU618675B2 (de)
CA (1)	CA2016211C (de)
DE (1)	DE69028040T2 (de)
DK (1)	DK0398539T3 (de)
ES (1)	ES2091224T3 (de)
GR (1)	GR3021010T3 (de)
SG (1)	SG47797A1 (de)
WO (1)	WO1990014420A1 (de)

Families Citing this family (43)

* Cited by examiner, † Cited by third party
Publication number	Priority date	Publication date	Assignee	Title
DE3856593D1 (de) *	1987-04-06	2007-10-04	Novozymes As	Regelung von elektrostatischen interaktionen an metallionen-Bindungsstellen zur Stabilisierung von Proteinen
US5665587A (en) *	1989-06-26	1997-09-09	Novo Nordisk A/S	Modified subtilisins and detergent compositions containing same
DK271490D0 (da) *	1990-11-14	1990-11-14	Novo Nordisk As	Detergentkomposition
US5863783A (en) *	1991-03-27	1999-01-26	Gist-Brocades, N.V.	Cloning and expression of DNA molecules encoding arabinan-degrading enzymes of fungal origin
CZ91194A3 (en) *	1991-10-16	1994-12-15	Unilever Nv	Stable aqueous enzymatic detergent and process for preparing thereof
US5989887A (en) *	1992-11-25	1999-11-23	Dsm N.V.	Cloning and expression of DNA molecules incoding arabinan-degrading enzymes of fungal origin
US5567601A (en) *	1993-06-01	1996-10-22	University Of Maryland	Subtilisin mutants lacking a primary calcium binding site
US6436690B1 (en)	1993-09-15	2002-08-20	The Procter & Gamble Company	BPN′ variants having decreased adsorption and increased hydrolysis wherein one or more loop regions are substituted
US6440717B1 (en)	1993-09-15	2002-08-27	The Procter & Gamble Company	BPN′ variants having decreased adsorption and increased hydrolysis
EP0723580B1 (de) *	1993-10-14	2003-07-16	The Procter & Gamble Company	Proteasen enthaltende bleichmittelzusammensetzungen
DE69434962T2 (de) *	1993-10-14	2008-01-17	The Procter & Gamble Company, Cincinnati	Proteasehaltige reinigungsmittel
MA23346A1 (fr) *	1993-10-14	1995-04-01	Genencor Int	Variantes de la subtilisine
DE4411223A1 (de) *	1994-03-31	1995-10-05	Solvay Enzymes Gmbh & Co Kg	Verwendung alkalischer Proteasen in gewerblichen Textilwaschverfahren
US6599730B1 (en)	1994-05-02	2003-07-29	Procter & Gamble Company	Subtilisin 309 variants having decreased adsorption and increased hydrolysis
US6066611A (en) *	1994-10-13	2000-05-23	The Procter & Gamble Company	Bleaching compositions comprising protease enzymes
US6455295B1 (en)	1995-03-08	2002-09-24	The Procter & Gamble Company	Subtilisin Carlsberg variants having decreased adsorption and increased hydrolysis
IL117350A0 (en)	1995-03-09	1996-07-23	Procter & Gamble	Proteinase k variants having decreased adsorption and increased hydrolysis
US6475765B1 (en)	1995-03-09	2002-11-05	Procter & Gamble Company	Subtilisin DY variants having decreased adsorption and increased hydrolysis
ES2193421T3 (es) *	1996-12-20	2003-11-01	Unilever Nv	Composicion de blanqueo enzimatica.
US6204234B1 (en) *	1997-07-09	2001-03-20	The Proctor & Gamble Company	Cleaning compositions comprising a specific oxygenase
AR015977A1 (es) *	1997-10-23	2001-05-30	Genencor Int	Variantes de proteasa multiplemente substituida con carga neta alterada para su empleo en detergentes
ES2368718T3 (es) *	1997-10-23	2011-11-21	Danisco Us Inc.	Variantes de subtilisina con múltiples sustituciones.
US6908757B1 (en)	1998-03-26	2005-06-21	The Procter & Gamble Company	Serine protease variants having amino acid deletions and substitutions
US6569663B1 (en)	1998-03-26	2003-05-27	The Procter & Gamble Company	Serine protease variants having amino acid substitutions
MXPA02000842A (es)	1999-07-22	2002-07-30	Procter & Gamble	Conjugados de proteasa que tienen sitios de corte protegidos estericamente.
CN1373802A (zh)	1999-07-22	2002-10-09	宝洁公司	在特定表位区域具有氨基酸缺失和取代的枯草杆菌蛋白酶变体
BR0012660A (pt)	1999-07-22	2002-04-09	Procter & Gamble	Variante de protease tipo subtilisina; composição de limpeza; e composição de cuidado pessoal
US6946128B1 (en)	1999-07-22	2005-09-20	The Procter & Gamble Company	Protease conjugates having sterically protected epitope regions
US7217554B2 (en)	1999-08-31	2007-05-15	Novozymes A/S	Proteases and variants thereof
WO2001016285A2 (en)	1999-08-31	2001-03-08	Novozymes A/S	Novel proteases and variants thereof
US6727085B2 (en) *	1999-12-15	2004-04-27	Fanoe Tina Sejersgaard	Subtilase variants having an improved wash performance on egg stains
US7087415B2 (en) *	2000-07-31	2006-08-08	Athena Biotechnologies, Inc.	Methods and compositions for directed gene assembly
WO2002069232A2 (en)	2001-02-19	2002-09-06	Merck Patent Gmbh	Method for identification of t-cell epitopes and use for preparing molecules with reeduced immunogenicity
PL210859B1 (pl) *	2001-03-23	2012-03-30	Genencor Int	Odmiana subtylizyny zawierająca zmieniony epitop limfocytów T, kwas nukleinowy kodujący tę odmianę subtylizyny, wektor ekspresyjny i komórka gospodarza zawierająca kwas nukleinowy oraz kompozycje zawierające tę odmianę subtylizyny
CA2455607C (en) *	2001-07-09	2013-01-08	Children's Hospital Medical Center Of Akron	Multiple controls for molecular genetic analyses
DE10153792A1 (de)	2001-10-31	2003-05-22	Henkel Kgaa	Neue Alkalische Protease-Varianten und Wasch- und Reinigungsmittel enthaltend diese neuen Alkalischen Protease-Varianten
EP2287320B1 (de) *	2002-01-16	2014-10-01	Danisco US Inc.	Mehrfach substituierte Proteasevarianten
US7888093B2 (en) *	2002-11-06	2011-02-15	Novozymes A/S	Subtilase variants
US7294499B2 (en)	2003-01-30	2007-11-13	Novozymes A/S	Subtilases
US7727756B2 (en) *	2003-03-21	2010-06-01	Novozymes A/S	Subtilases
JP4819670B2 (ja) *	2003-05-07	2011-11-24	ノボザイムスアクティーゼルスカブ	変異体スブチリシン酵素（サブチラーゼ）
US12509672B2 (en)	2018-11-28	2025-12-30	Danisco Us Inc.	Subtilisin variants having improved stability
EP4525615A2 (de)	2022-05-14	2025-03-26	Novozymes A/S	Zusammensetzungen und verfahren zur prävention, behandlung, unterdrückung und/oder beseitigung von phytopathogenen befällen und infektionen

Family Cites Families (12)

* Cited by examiner, † Cited by third party
Publication number	Priority date	Publication date	Assignee	Title
US4760025A (en) *	1984-05-29	1988-07-26	Genencor, Inc.	Modified enzymes and methods for making same
US5155033A (en) *	1989-01-06	1992-10-13	Genencor, Inc.	Subtilisins modified at position 225 resulting in a shift in catalytic activity
EP0254735B2 (de) *	1986-01-15	1998-06-17	Amgen Inc.	Herstellung von ph- und wärmestabile subtilisinanaloge
US5013657A (en) *	1988-04-12	1991-05-07	Bryan Philip N	Subtilisin mutations
US4990452A (en) *	1986-02-12	1991-02-05	Genex Corporation	Combining mutations for stabilization of subtilisin
US4980288A (en) *	1986-02-12	1990-12-25	Genex Corporation	Subtilisin with increased thermal stability
IE65767B1 (en) *	1986-04-30	1995-11-15	Genencor Int	Non-human carbonyl hydrolase mutants DNA sequences and vectors encoding same and hosts transformed with said vectors
NZ221627A (en) *	1986-09-09	1993-04-28	Genencor Inc	Preparation of enzymes, modifications, catalytic triads to alter ratios or transesterification/hydrolysis ratios
ES2135386T3 (es) *	1987-02-27	1999-11-01	Genencor Int	Transformacion de cepas de bacillus alcalofilas.
DE3856593D1 (de) *	1987-04-06	2007-10-04	Novozymes As	Regelung von elektrostatischen interaktionen an metallionen-Bindungsstellen zur Stabilisierung von Proteinen
US4914031A (en) *	1987-04-10	1990-04-03	Amgen, Inc.	Subtilisin analogs
FR2614467A1 (fr) *	1987-04-21	1988-10-28	Telemecanique Electrique	Appareil interrupteur de protection a mecanisme de declenchement simplifie

1990
- 1990-04-23 KR KR1019910700067A patent/KR100188532B1/ko not_active Expired - Fee Related
- 1990-04-23 WO PCT/US1990/002234 patent/WO1990014420A1/en not_active Ceased
- 1990-04-23 JP JP2507107A patent/JPH03505976A/ja active Pending
- 1990-04-23 AU AU55580/90A patent/AU618675B2/en not_active Expired
- 1990-05-01 DE DE69028040T patent/DE69028040T2/de not_active Expired - Lifetime
- 1990-05-01 EP EP90304715A patent/EP0398539B1/de not_active Expired - Lifetime
- 1990-05-01 SG SG1996004414A patent/SG47797A1/en unknown
- 1990-05-01 ES ES90304715T patent/ES2091224T3/es not_active Expired - Lifetime
- 1990-05-01 AT AT90304715T patent/ATE141324T1/de not_active IP Right Cessation
- 1990-05-01 DK DK90304715.7T patent/DK0398539T3/da active
- 1990-05-07 CA CA002016211A patent/CA2016211C/en not_active Expired - Lifetime
1992
- 1992-03-25 US US07/857,714 patent/US5397705A/en not_active Expired - Lifetime
1996
- 1996-09-12 GR GR960402360T patent/GR3021010T3/el unknown

Also Published As

Publication number	Publication date
SG47797A1 (en)	1998-04-17
HK1006857A1 (en)	1999-03-19
EP0398539A1 (de)	1990-11-22
DE69028040T2 (de)	1997-02-27
JPH03505976A (ja)	1991-12-26
ATE141324T1 (de)	1996-08-15
DE69028040D1 (de)	1996-09-19
US5397705A (en)	1995-03-14
KR100188532B1 (ko)	1999-06-01
AU5558090A (en)	1990-12-18
CA2016211A1 (en)	1990-11-17
GR3021010T3 (en)	1996-12-31
ES2091224T3 (es)	1996-11-01
KR920701426A (ko)	1992-08-11
AU618675B2 (en)	1992-01-02
WO1990014420A1 (en)	1990-11-29
EP0398539B1 (de)	1996-08-14
DK0398539T3 (da)	1996-12-09

Legal Events

Date	Code	Title	Description
1990-05-07	EEER	Examination request
2010-05-07	MKEX	Expiry

Publication	Publication Date	Title
CA2016211C (en)	1998-07-21	Multiply mutated subtilisins
EP0309565B2 (de)	2001-08-22	Subtilisin-analoge
US5399283A (en)	1995-03-21	Thermally stable and pH stable subtilisin analogs and method for production thereof
EP0328229B1 (de)	1994-01-19	Proteolytische Enzyme und ihre Verwendung in Waschmitteln
AU604476C (en)	1993-08-12	Thermally stable and pH stable subtilisin analogs and method for production thereof
HK1006857B (en)	1999-03-19	Multiply mutated subtilisins
HK1007168B (en)	1999-04-01	Subtilisins analogs
NO176844B (no)	1995-02-27	Enzymatisk aktive analoger av et Bacillus-subtilisin, nukleinsyre, vertscelle, fremgangsmåte for forbedring av varme- og pH-stabiliteten til et slikt subtilisin, samt vaskemiddelblanding som omfatter en slik analog